Objective:To explore the relationship between sperm DNA integrity, semen parameters, seminal plasma neutral α-glucosidase activity and in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcomes. Methods:The clinical data of 300 male infertile patients who underwent routine semen analysis in the Second Affiliated Hospital of Air Force Medical University from October 2023 to April 2024 and whose spouses received IVF/ICSI treatment were retrospectively analyzed. The results of sperm DNA integrity [sperm DNA fragmentation index (DFI)], semen parameters (sperm percentage, sperm density, sperm motility, normal morphology sperm rate) and seminal plasma neutral α-glucosidase activity at admission were recorded. According to their spouses′ IVF/ICSI outcomes, the 300 patients were divided into the successful pregnancy group (those with successful intrauterine pregnancy detected by ultrasound 30 days after transplantation) and the unsuccessful pregnancy group (those without successful intrauterine pregnancy detected by ultrasound 30 days after transplantation). The differences in sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity at admission between the two groups were compared. The receiver operating characteristic (ROC) curve was used to analyze the predictive efficacy of sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity in male infertile patients at admission for their spouses′ IVF/ICSI outcomes.Results:According to the IVF/ICSI outcomes of the spouses of male infertile patients, 169 cases (56.3%) had successful intrauterine pregnancy detected by ultrasound 30 days after transplantation; 131 cases (43.7%) had unsuccessful intrauterine pregnancy. At admission, there was no statistically significant difference in clinical data between the two groups (all P>0.05). The DFI and sperm density in the successful pregnancy group were lower than those in the unsuccessful pregnancy group, while the sperm percentage, sperm motility, normal morphology sperm rate and seminal plasma neutral α-glucosidase activity were higher than those in the unsuccessful pregnancy group (all P<0.05). The area under the ROC curve (AUC) values of single indicators including DFI, sperm percentage, sperm density, sperm motility, normal morphology sperm rate, and seminal plasma neutral α-glucosidase level in predicting the IVF/ICSI outcomes of spouses of male infertile patients were 0.719, 0.718, 0.812, 0.779, 0.769, and 0.736, respectively; the sensitivities were 70.42%, 77.46%, 69.01%, 70.42%, 69.01%, and 77.46%, respectively; the specificities were 66.38%, 55.02%, 81.22%, 73.80%, 77.29%, and 62.88%, respectively; the Youden indexes were 0.368, 0.325, 0.502, 0.442, 0.463, and 0.404, respectively; all differences were statistically significant (all P<0.05). Conclusions:Sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity can assist in predicting IVF/ICSI outcomes, providing an important reference for the treatment outcomes of male infertile patients.

Objective:To investigate the characteristic distribution of vaginal microbiota in infertile women.Methods:We collected the results of vaginal microbiological examinations from 19322 initial visit infertile women who visited the Reproductive Medicine Center of the Department of Obstetrics and Gynecology,the Second Affili-ated Hospital of Air Force Medical University from March 1,2023 to July 31,2024.The vaginal microbiota infection status of patients was compared in different age groups(＜25 years old,25-＜30 years old,30-＜35 years old,35-＜40 years old,≥40 years old)and different seasons(spring,summer,autumn,winter).Results:①Among 19322 women,6027 cases(31.19％)showed abnormal vaginal microecology.Pathogenic microorganisms were detected in 3093 cases,including 2882 cases of single vaginitis,211 cases of mixed vaginitis,3764 cases with vagi-nal cleanliness grade Ⅲ-Ⅳ,and 3965 cases with abnormal lactobacilli.Among patients with single vaginitis,1349 cases(46.81％)were diagnosed with vulvovaginal candidiasis(VVC),which was the highest proportion.Aerobic vaginitis(AV)followed with 752 cases(26.09％),and bacterial vaginosis(BV)had 671 cases(23.28％),trichomonal vaginitis(TV)with 110 cases(3.82％).Among patients with mixed vaginitis,AV+BV was the most common with 96 cases(45.49％).②The detection rates of cleanliness grade Ⅲ-Ⅳ,abnormal lactobacilli,abnor-mal microorganisms(unclear pathogen),single vaginitis(BV,VVC and TV),and mixed vaginitis showed statisti-cally significant differences across different seasons(P＜0.05).Specifically,the detection rates of cleanliness grade Ⅲ-Ⅳ and abnormal microorganisms(unclear pathogens)were significantly higher in autumn than in other seasons(P＜0.05),while the detection rate of abnormal lactobacilli was higher in spring than in other seasons(P＜0.05).③The detection rates of abnormal lactobacilli,abnormal microorganisms(unclear pathogen),single vaginitis(BV,VVC and AV),and mixed vaginitis showed significant differences among different age groups(P＜0.05).Specifically,the detection rate of abnormal microorganisms(unclear pathogen)was higher in the age group＜25 years than in other age groups(P＜0.05),while the detection rate of BV among single vaginitis cases was higher in the age group≥40 years than in other age groups(P＜0.05).Conclusions:The vaginal microecol-ogy of infertile women varies in terms of infection rates across different age groups and seasons.Patients with simple vaginitis have the highest rate of VVC,while those with mixed vaginitis have the highest proportion of AV+BV infection.

Background and purpose:Next generation sequencing-identified genetic alterations of diffuse large B cell lymphoma(DLBCL)and baseline SUVmax detected by 18F-FDG PET/CT were correlated with patients'prognosis.However,their relationship and the associations with R-CHOP response of DLBCL are still unclear.This study aimed to analyze the association bewteen genetic alterations and 18F-FDG PET/CT SUVmax and their correlations with clinicopathological characteristics and R-CHOP response of DLBCL.Methods:A total of 225 cases of primary DLBCL detected by next generation sequencing using 481 lymphoma gene panel and examined by 18F-FDG PET/CT before treatment between 2022 and 2023 were collected.This study was approved by the Ethics Committee of Fudan University Shanghai Cancer Center(Ethical No.:050432-4-2307E)and acquired the informed consent of the patients.The translocations of BCL2,BCL6 and MYC were identified by fluorescence in situ hybridization.The clinicopathological characteristics and the PET/CT scan after R-CHOP chemotherapy were collected.Results:Finally,191 patients were enrolled in this study.The frequency of MYD88 mutation,TP53 mutation,copy number variations of CDKN2A/2B,CD79B mutation in the 191 DLBCL patients were 24.6%,27.2%,32.5%and 16.8%,respectively.The range of baseline SUVmax was 5.10-63.10(24.44±10.70,median 22.80).The baseline SUVmax of MYD88L265P DLBCL was significantly higher than that of MYD88 wild type(P=0.039).There were no significant associations of SUVmax with other gene alterations including TP53 mutation,CDKN2A/B loss,CD79B mutation,KMT2D mutation,TNFAIP3 mutation,B2M mutation,EZH2 mutation,BTG1/2 mutation,CREBBP mutation,gene translocations of MYC,BCL2 and BCL6.The higher SUVmax before treatment was correlated with higher serum lactate dehydrogenase(LDH)level(P=0.012)and non-germinal center B-cell-like(non-GCB)DLBCL(P=0.040).However,there was no significant association of SUVmax with R-CHOP response(P=0.714).TP53 mutation was significantly associated with the poor response of R-CHOP(P=0.001)and was an independent predictor of non-complete metabolic response(non-CMR).TP53 mutation combined with Ann Arbor stage,International Prognostic Index(IPI)score and serum LDH level could better predict R-CHOP response than each factor alone.Conclusion:MYD88L265P DLBCL had higher baseline 18F-FDG PET/CT SUVmax.The baseline SUVmax was not associated with R-CHOP response.However,TP53 mutation was significantly correlated with poor response of R-CHOP in DLBCL patients.TP53 mutation combined with clinicopathological characteristics could better predict R-CHOP response.The associations of gene alterations and SUVmax with prognosis of DLBCL patients needed to be explored in the future.

Objective To explore the clinical effect of applying an improved"outside-in"technique in hip arthroscopic surgery.Methods Totally 136 patients undergoing hip arthroscopic surgery between June 2021 and July 2023 were selected and studied retrospectively.According to their different surgi-cal approach,they were divided into a modified approach group(n=75,including 30 males and 45 fe-males,with an average age of 36±14)and a classic approach group(n=61,including 33 males and 28 females,with an average age of 31±11).Before as well as 4 weeks,3 months and 6 months after the surgery,both groups were evaluated using the visual analog scale(VAS)and Harris hip score,and observed their surgical complications.Moreover,the surgical outcomes and time of sur-gical approach establishment were compared between the two groups.Results There was no significant difference between the two groups in general information,preoperative VAS and Harris scores.More-over,no significant difference was found between the two groups in the Harris score after surgery.Compared with the classic approach group,the improved group had significantly less surgical time(49.0±16.9 min vs.66.0±13.3 min,P<0.05),without using the C-arm fluoroscopy during surgery.Moreover,in the modified approach group,the time for establishing the mid-anteriorapproach was sig-nificantly shortened(10.4±5.9 min vs.25.9±15.1 min,P<0.05),along with the traction time during surgery(66.0±13.3 min vs.49.0±16.9 min,P<0.05).Conclusion The modified"outside-in"hip ar-throscopy technique is a safe and effective surgical method,easier to operate,with shorter surgical time.

Objective:To investigate the characteristic distribution of vaginal microbiota in infertile women.Methods:We collected the results of vaginal microbiological examinations from 19322 initial visit infertile women who visited the Reproductive Medicine Center of the Department of Obstetrics and Gynecology,the Second Affili-ated Hospital of Air Force Medical University from March 1,2023 to July 31,2024.The vaginal microbiota infection status of patients was compared in different age groups(＜25 years old,25-＜30 years old,30-＜35 years old,35-＜40 years old,≥40 years old)and different seasons(spring,summer,autumn,winter).Results:①Among 19322 women,6027 cases(31.19％)showed abnormal vaginal microecology.Pathogenic microorganisms were detected in 3093 cases,including 2882 cases of single vaginitis,211 cases of mixed vaginitis,3764 cases with vagi-nal cleanliness grade Ⅲ-Ⅳ,and 3965 cases with abnormal lactobacilli.Among patients with single vaginitis,1349 cases(46.81％)were diagnosed with vulvovaginal candidiasis(VVC),which was the highest proportion.Aerobic vaginitis(AV)followed with 752 cases(26.09％),and bacterial vaginosis(BV)had 671 cases(23.28％),trichomonal vaginitis(TV)with 110 cases(3.82％).Among patients with mixed vaginitis,AV+BV was the most common with 96 cases(45.49％).②The detection rates of cleanliness grade Ⅲ-Ⅳ,abnormal lactobacilli,abnor-mal microorganisms(unclear pathogen),single vaginitis(BV,VVC and TV),and mixed vaginitis showed statisti-cally significant differences across different seasons(P＜0.05).Specifically,the detection rates of cleanliness grade Ⅲ-Ⅳ and abnormal microorganisms(unclear pathogens)were significantly higher in autumn than in other seasons(P＜0.05),while the detection rate of abnormal lactobacilli was higher in spring than in other seasons(P＜0.05).③The detection rates of abnormal lactobacilli,abnormal microorganisms(unclear pathogen),single vaginitis(BV,VVC and AV),and mixed vaginitis showed significant differences among different age groups(P＜0.05).Specifically,the detection rate of abnormal microorganisms(unclear pathogen)was higher in the age group＜25 years than in other age groups(P＜0.05),while the detection rate of BV among single vaginitis cases was higher in the age group≥40 years than in other age groups(P＜0.05).Conclusions:The vaginal microecol-ogy of infertile women varies in terms of infection rates across different age groups and seasons.Patients with simple vaginitis have the highest rate of VVC,while those with mixed vaginitis have the highest proportion of AV+BV infection.

Objective To explore the clinical effect of applying an improved"outside-in"technique in hip arthroscopic surgery.Methods Totally 136 patients undergoing hip arthroscopic surgery between June 2021 and July 2023 were selected and studied retrospectively.According to their different surgi-cal approach,they were divided into a modified approach group(n=75,including 30 males and 45 fe-males,with an average age of 36±14)and a classic approach group(n=61,including 33 males and 28 females,with an average age of 31±11).Before as well as 4 weeks,3 months and 6 months after the surgery,both groups were evaluated using the visual analog scale(VAS)and Harris hip score,and observed their surgical complications.Moreover,the surgical outcomes and time of sur-gical approach establishment were compared between the two groups.Results There was no significant difference between the two groups in general information,preoperative VAS and Harris scores.More-over,no significant difference was found between the two groups in the Harris score after surgery.Compared with the classic approach group,the improved group had significantly less surgical time(49.0±16.9 min vs.66.0±13.3 min,P<0.05),without using the C-arm fluoroscopy during surgery.Moreover,in the modified approach group,the time for establishing the mid-anteriorapproach was sig-nificantly shortened(10.4±5.9 min vs.25.9±15.1 min,P<0.05),along with the traction time during surgery(66.0±13.3 min vs.49.0±16.9 min,P<0.05).Conclusion The modified"outside-in"hip ar-throscopy technique is a safe and effective surgical method,easier to operate,with shorter surgical time.

Background and purpose:Next generation sequencing-identified genetic alterations of diffuse large B cell lymphoma(DLBCL)and baseline SUVmax detected by 18F-FDG PET/CT were correlated with patients'prognosis.However,their relationship and the associations with R-CHOP response of DLBCL are still unclear.This study aimed to analyze the association bewteen genetic alterations and 18F-FDG PET/CT SUVmax and their correlations with clinicopathological characteristics and R-CHOP response of DLBCL.Methods:A total of 225 cases of primary DLBCL detected by next generation sequencing using 481 lymphoma gene panel and examined by 18F-FDG PET/CT before treatment between 2022 and 2023 were collected.This study was approved by the Ethics Committee of Fudan University Shanghai Cancer Center(Ethical No.:050432-4-2307E)and acquired the informed consent of the patients.The translocations of BCL2,BCL6 and MYC were identified by fluorescence in situ hybridization.The clinicopathological characteristics and the PET/CT scan after R-CHOP chemotherapy were collected.Results:Finally,191 patients were enrolled in this study.The frequency of MYD88 mutation,TP53 mutation,copy number variations of CDKN2A/2B,CD79B mutation in the 191 DLBCL patients were 24.6%,27.2%,32.5%and 16.8%,respectively.The range of baseline SUVmax was 5.10-63.10(24.44±10.70,median 22.80).The baseline SUVmax of MYD88L265P DLBCL was significantly higher than that of MYD88 wild type(P=0.039).There were no significant associations of SUVmax with other gene alterations including TP53 mutation,CDKN2A/B loss,CD79B mutation,KMT2D mutation,TNFAIP3 mutation,B2M mutation,EZH2 mutation,BTG1/2 mutation,CREBBP mutation,gene translocations of MYC,BCL2 and BCL6.The higher SUVmax before treatment was correlated with higher serum lactate dehydrogenase(LDH)level(P=0.012)and non-germinal center B-cell-like(non-GCB)DLBCL(P=0.040).However,there was no significant association of SUVmax with R-CHOP response(P=0.714).TP53 mutation was significantly associated with the poor response of R-CHOP(P=0.001)and was an independent predictor of non-complete metabolic response(non-CMR).TP53 mutation combined with Ann Arbor stage,International Prognostic Index(IPI)score and serum LDH level could better predict R-CHOP response than each factor alone.Conclusion:MYD88L265P DLBCL had higher baseline 18F-FDG PET/CT SUVmax.The baseline SUVmax was not associated with R-CHOP response.However,TP53 mutation was significantly correlated with poor response of R-CHOP in DLBCL patients.TP53 mutation combined with clinicopathological characteristics could better predict R-CHOP response.The associations of gene alterations and SUVmax with prognosis of DLBCL patients needed to be explored in the future.

Objective:To explore the relationship between sperm DNA integrity, semen parameters, seminal plasma neutral α-glucosidase activity and in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcomes. Methods:The clinical data of 300 male infertile patients who underwent routine semen analysis in the Second Affiliated Hospital of Air Force Medical University from October 2023 to April 2024 and whose spouses received IVF/ICSI treatment were retrospectively analyzed. The results of sperm DNA integrity [sperm DNA fragmentation index (DFI)], semen parameters (sperm percentage, sperm density, sperm motility, normal morphology sperm rate) and seminal plasma neutral α-glucosidase activity at admission were recorded. According to their spouses′ IVF/ICSI outcomes, the 300 patients were divided into the successful pregnancy group (those with successful intrauterine pregnancy detected by ultrasound 30 days after transplantation) and the unsuccessful pregnancy group (those without successful intrauterine pregnancy detected by ultrasound 30 days after transplantation). The differences in sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity at admission between the two groups were compared. The receiver operating characteristic (ROC) curve was used to analyze the predictive efficacy of sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity in male infertile patients at admission for their spouses′ IVF/ICSI outcomes.Results:According to the IVF/ICSI outcomes of the spouses of male infertile patients, 169 cases (56.3%) had successful intrauterine pregnancy detected by ultrasound 30 days after transplantation; 131 cases (43.7%) had unsuccessful intrauterine pregnancy. At admission, there was no statistically significant difference in clinical data between the two groups (all P>0.05). The DFI and sperm density in the successful pregnancy group were lower than those in the unsuccessful pregnancy group, while the sperm percentage, sperm motility, normal morphology sperm rate and seminal plasma neutral α-glucosidase activity were higher than those in the unsuccessful pregnancy group (all P<0.05). The area under the ROC curve (AUC) values of single indicators including DFI, sperm percentage, sperm density, sperm motility, normal morphology sperm rate, and seminal plasma neutral α-glucosidase level in predicting the IVF/ICSI outcomes of spouses of male infertile patients were 0.719, 0.718, 0.812, 0.779, 0.769, and 0.736, respectively; the sensitivities were 70.42%, 77.46%, 69.01%, 70.42%, 69.01%, and 77.46%, respectively; the specificities were 66.38%, 55.02%, 81.22%, 73.80%, 77.29%, and 62.88%, respectively; the Youden indexes were 0.368, 0.325, 0.502, 0.442, 0.463, and 0.404, respectively; all differences were statistically significant (all P<0.05). Conclusions:Sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity can assist in predicting IVF/ICSI outcomes, providing an important reference for the treatment outcomes of male infertile patients.

BACKGROUND:De novo adipose regeneration induced by small extracellular vesicles has become a promising method for repairing soft tissue defects.However,due to different animal models and small extracellular vesicle application dosages,it is difficult to quantitatively compare the therapeutic effect of small extracellular vesicles from various sources on adipose regeneration. OBJECTIVE:To compare the regenerative effects of small extracellular vesicles derived from stem cells and small extracellular vesicles from tissue. METHODS:Small extracellular vesicles derived from adipose-derived stem cells and from adipose tissue were isolated by ultracentrifugation.The particle number,particle size,morphology,and protein expression of small extracellular vesicles were identified by nanoparticle tracking analysis,transmission electron microscopy and western blot assay.A quantitative and evaluative subcutaneous model for adipose regeneration in C57 mice was established using a customized silicone tube.The regenerative effects of induced de novo adipose were compared by cell counting and hematoxylin-eosin staining. RESULTS AND CONCLUSION:(1)Small extracellular vesicles derived from adipose-derived stem cells and from adipose tissue were isolated by ultracentrifugation.Both small extracellular vesicles were round-shape in transmission electron microscopy with particle size between 50-200 nm,and abundant with the small extracellular vesicles marker protein CD81,CD63 and TSG101.(2)An equal number of small extracellular vesicles were mixed with matrigel in customized silicone tubes,implanted subcutaneously in the back of mice to establish a cell-free and quantifiable adipose regeneration model.(3)On days 3 and 7 after implantation,the results of cell counting and hematoxylin-eosin staining showed that both small extracellular vesicle groups recruited more host cells than the blank group,and the small extracellular vesicles derived from adipose tissue group were superior to the small extracellular vesicles derived from adipose-derived stem cell group.(4)4 weeks after implantation,hematoxylin-eosin staining of the contents in silicone tubes showed that small extracellular vesicles induced de novo adipose regeneration in vivo,and the small extracellular vesicles derived from adipose tissue group were superior to the small extracellular vesicles derived from adipose-derived stem cell group.The above results indicated that small extracellular vesicles derived from tissues have a superior effect on inducing de novo adipose regeneration compared to small extracellular vesicles derived from stem cells.

[Abstract] [Objective] To construct inducible immortalization gene vectors for transfection into primary cells, enabling the establishment of a conditionally immortalized cell line that support their sustained cultivation and proliferation in vitro. [Methods] Using gene homologous recombination technology, the coding sequences (CDS) of immortalization genes-including human telomerase reverse transcriptase (hTERT), simian virus 40 large T antigen (SV40LT), acute myeloid leukemia fusion genes NUP98-KDM5A (N/K) and CBFA2T3-GLIS2 (C/G), as well as the proto-oncogene KRAS were precisely inserted into the tetracycline (Tet)-inducible eukaryotic expression lentiviral vector pLV2-TRE3GS-EGFP-MCS-3×FLAG-hPGK-Tet-On-SV40-Neo and the transposon PB-TRE3G-3×FLAG-T2A-Puro-SV40-PA. Lentiviral packaging, cell transfection, mRNA expression analysis, Western blotting for protein detection, green fluorescent protein (GFP) visualization, and cell proliferation assays were conducted to evaluate transfection efficiency and assess the regulatory effects of Tet on gene expression in 293T and MEF cells. [Results] The Tet-inducible lentiviral vectors pLV2-Tet-SV40LT, pLV2-Tet-N/K, and pLV2-Tet-C/G, along with the transposon vectors PB-Tet-hTERT, PB-Tet-SV40LT, PB-Tet-N/K, PB-Tet-C/G, and PB-Tet-KRAS, were successfully constructed. In 293T cells, the expression levels of all target genes were upregulated after transfection. In MEF cells, the immortalizing functions of SV40LT and N/K were validated. By modulating Tet addition, cell proliferation levels were effectively regulated, leading to the successful establishment of conditionally immortalized pLV2-SV40LT-MEF and pLV2-N/K-MEF cell lines. [Conclusion] The construction of Tet-inducible immortalizing gene vectors provides a technical foundation for establishing conditionally immortalized primary cell lines, thereby facilitating research on the large-scale in vitro production and expansion of blood cells, such as erythrocytes and platelets.